title: PK modulation of haptenylated peptides via non-covalent antibody complexation.
abstract: We applied noncovalent complexes of digoxigenin (Dig) binding antibodies with digoxigeninylated peptide derivatives to modulate their pharmacokinetic properties. A peptide derivative which activates the Y2R receptor was selectively mono-digoxigeninylated by reacting a NHS-Dig derivative with an ε-amino group of lysine 2. This position tolerates modifications without destroying receptor binding and functionality of the peptide. Dig-peptide derivatives can be loaded onto Dig-binding IgGs in a simple and robust reaction, thereby generating peptide-IgG complexes in a defined two to one molar ratio. This indicates that each antibody arm becomes occupied by one haptenylated peptide. In vitro receptor binding and signaling assays showed that Dig-peptides as well as the peptide-antibody complexes retain better potency than the corresponding pegylated peptides. In vivo analyses revealed prolonged serum half-life of antibody-complexed peptides compared to unmodified peptides. Thus, complexes are of sufficient stability for PK modulation. We observed more prolonged weight reduction in a murine diet-induced obesity (DIO) model with antibody-complexed peptides compared to unmodified peptides. We conclude that antibody-hapten complexation can be applied to modulate the PK of haptenylated peptides and in consequence improve the therapeutic efficacy of therapeutic peptides.
authors.affiliation: Roche Pharma Research & Early Development pRED, Large Molecule Research, Nonnenwald 2, D-82372 Penzberg, Germany. eike.hoffmann@roche.com
authors.forename: Eike
authors.initials: E
authors.lastname: Hoffmann
authors.forename: Anish
authors.initials: A
authors.lastname: Konkar
authors.forename: Sebastian
authors.initials: S
authors.lastname: Dziadek
authors.forename: Hans-Peter
authors.initials: HP
authors.lastname: Josel
authors.forename: Karin
authors.initials: K
authors.lastname: Conde-Knape
authors.forename: Holger
authors.initials: H
authors.lastname: Kropp
authors.forename: Lothar
authors.initials: L
authors.lastname: Kling
authors.forename: Kay
authors.initials: K
authors.lastname: Stubenrauch
authors.forename: Irmgard
authors.initials: I
authors.lastname: Thorey
authors.forename: Stefan
authors.initials: S
authors.lastname: Dengl
authors.forename: Ulrich
authors.initials: U
authors.lastname: Brinkmann
mesh_terms: D000818:Animals; D059305:Diet, High-Fat; D004076:Digoxigenin / Q000097:blood / Q000737:chemistry* / Q000493:pharmacokinetics; D004435:Eating / Q000187:drug effects; D005260:Female; D057809:HEK293 Cells; D006241:Haptens / Q000737:chemistry*; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D009765:Obesity / Q000188:drug therapy; D010455:Peptides / Q000737:chemistry* / Q000493:pharmacokinetics; D058912:Purinergic P2Y Receptor Agonists / Q000008:administration & dosage; D058470:Receptors, Purinergic P2Y / Q000378:metabolism
chemical_list: D006241:Haptens; D007074:Immunoglobulin G; D010455:Peptides; D058912:Purinergic P2Y Receptor Agonists; D058470:Receptors, Purinergic P2Y; D004076:Digoxigenin
keywords: CME; Diabetes; Dig; Digoxigenin; H-chain; L-chain; PYY; Pharmacokinetics; SEC; SPR; carboxymethyl ether; digoxigenin; heavy chain; light chain; size exclusion chromatography; surface plasmon resonance